NZ526642A

NZ526642A - Process for preparing aqueous extracts of plants with the additional treatment step cpmprising treating the comminuted plant with laser radiation and the plant extracts so obtained

Info

Publication number: NZ526642A
Application number: NZ526642A
Authority: NZ
Inventors: Pena Jose Manuel Frias
Original assignee: Bomsund Grupo Asesor Sl
Priority date: 2000-11-27
Filing date: 2000-11-27
Publication date: 2004-11-26
Also published as: KR20030063393A; BR0017378A; US7402324B1; MXPA03004721A; WO2002041908A1; DK1339420T3; HUP0501092A2; HRP20030482A2; CA2430113A1; NO20032369D0; CN1261162C; ES2228641T3; MY129109A; CN1479624A; NO20032369L; EP1339420B1; EP1339420A1; DE60013919D1; UA75624C2; PT1339420E

Abstract

A method for preparing aqueous extracts of vegetables, particularly of plants is described, which comprises the steps of a) decontamination of the plant, b) comminuting the plant, c) treatment of the comminuted plant with a laser radiation, d) suspension of the mixture obtained in step c) in water, e) maceration of the suspension obtained in step d), and f) separation of the resulting liquid. Compositions obtained by the present method are also described, some of which find application in medicine, particularly in the treatment of immune-suppressant diseases such as cancer, tuberculosis, influenza, common cold and AIDS, or in the treatment of viral diseases such as hepatitis.

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number 526642 526642 WO 02/41908 PCT/IB00/01947 1 PROCESS FOR PREPARING AQUEOUS EXTRACTS OF PLANTS AND EXTRACTS SO OBTAINED FIELD OF THE INVENTION The present invention relates to a process for preparing aqueous extracts of plants and to the aqueous extracts thus obtained, some of which find application in medicine, particularly in the treatment of immune suppressant diseases or in the treatment of viral diseases. BACKGROUND OF THE INVENTION Plants and, in general, vegetals are still an important source of active compositions and compounds employed in medicine. New plants, new compounds present in plants or even modifications of compounds present in plants are of potential interest to the phitochemical industry. The methods employed in the extraction of the components of vegetals and plants are of great importance regarding not only the yields of the compounds to be obtained, but also the chemical nature of the compounds which can in fact be gained. The usual methods employed in the phitochemical industry are based on the extraction of the comminuted plant or vegetal with water or steam, with organic solvents, or with mixtures of water and organic solvents such as water and alcohols. The inclusion of a maceration step is also well known in the art. Finally, the separation of the aqueous extract from the solid phase can be effected by conventional methods such as decantation, centrifiigation or filtration. Additional chemical treatments to modify the structure of the compounds present in the plants can also be effected. For more information on the subject, see Dr. Pio Font Quer, "Medicinal Plants. The updated Discorides" Ed. Labor, S.A., 12th Edition, 1990. The French patent FR 2733419 describes the preparation of extracts of a mixture of at least three different plants selected from the genus Geranium, Plantago and Calendula by steeping 300-400 g of fresh Geranium robertanium leaves, 10-50 g of dried Plantago lanceolata leaves and 10-50 g dried Calendula officinalis leaves in 3 1. of water at 15-30 °C for 3-15 days. According to FR 2733419, the composition thus obtained finds application in the treatment of cancer. However, a non- WO 02/41908 2 PCT/IB00/01947 satisfactory activity has been showed by these extracts, as reflected hi the examples below. The European patent application EP 0 934 746 A discloses the preparation of an aqueous extract of plants from the genus Geranium and Plantago as well as from the species Calendula officinalis, wherein said extracts are obtained from the following amounts in grams of said plants in water: 10 to 60 of dried Geranium sibiricun or 300 to 360 of fresh Geranium sibiricum, 10 to 60 of dried Plantago lanceolata, and 10 to 60 of dried Calendula Officinalis in 300 grams of water. The method for producing said aqueous extracts comprises macerating the comminuted plants in water before press-filtering the mixture and submitting it to a centrifugal treatment. According to EP 0 934 746 A, this extract may be used in oncological practice as part of a cancer therapeutic program. However, non-satisfactory activity has been showed by these extracts as reflected in the examples below. There is, thus, a need in the art for methods for preparing alternative extracts of plants which can be used as therapeutic agents, and which eventually can provide access to new compounds. SUMMARY OF THE INVENTION The present invention provides a method for preparing aqueous extracts of vegetals, particularly of plants, as well as the compositions obtained by this method, some of which find application in medicine, particularly in the treatment of immune-suppressant diseases such as cancer, tuberculosis, influenza, common cold, allergies, lupus erythematosus, psoriasis and AIDS, or in the treatment of viral diseases such as hepatitis. An aspect of the invention relates thus to a method for preparing aqueous extracts of vegetals, particularly of plants, which comprises the following steps: a) Decontamination of the plant b) Comminuting the plant. c) Treatment of the comminuted plant with a laser radiation. d) Suspension of the mixture obtained in step c) in water. e) Maceration of the suspension obtained in step d). WO 02/41908 3 PCT/IB00/01947 f) Separation of the resulting liquid. A second aspect of the invention relates to the compositions or aqueous extracts (hydrolates) obtained by the present method. A further aspect of the invention relates to the use of the present composition as therapeutic agent in the treatment of immune suppressant diseases such as cancer, tuberculosis, influenza, common cold, allergies, lupus erythematosus, psoriasis and AIDS; or in the treatment of viral diseases such as hepatitis. A further aspect of the invention relates to pharmaceutical compositions comprising the aqueous extracts obtained by the present method. The method of the invention differs from those of the cited prior art in the fact that the comminuted plant is treated with a laser radiation. As demonstrated in the examples below, the treatment of the comminuted plants with the laser results in either the presence of new activities or in unexpectedly high increments of the activities of the extracts. Therefore, the extracts thus obtained have necessarily to be different from those obtained by methods which do not effect the laser treatment. DETAILED DESCRIPTION OF THE INVENTION As mentioned above, the first aspect of the invention relates to a method for preparing aqueous extracts of vegetals, particularly of plants, which comprises the steps of: a) Decontamination of the plant b) Comminuting the plant. c) Treatment of the comminuted plant with a laser radiation. d) Suspension of the mixture obtained in step c) in water. e) Maceration of the suspension obtained in step d). f) Separation of the resulting liquid. WO 02/41908 4 PCT/IB00/01947 By the term "plant" it is understood either a unit or several units of the same' species, or several units of different species, or part or parts of a plant such as stems, leaves, flowers, etc. The decontamination (step a) is effected by washing the plant with water. The amount of water employed in this step is not determinant, and can be varied depending on the contamination state of the plant. Although higher and lower temperatures are not discarded, the water temperature should be between 10 and 40 °C, preferably between 20 and 35°C, and most preferably 28°C. A washing tunnel can be employed to facilitate this step. Both the amount of water and the residence time of the plant in the washing tunnel are not determinant, and can therefore be varied depending on the contamination state of the plant. The washing step can be carried out several times, with a drying step in between This drying step is preferably effected by placing the plant in the sun. Once the plant has been thoroughly decontaminated, it is comminuted (step b) by conventional methods such as a comminuting machine or even manually. Although higher and lower temperatures are not discarded, the temperature at which the plant is comminuted should be between 10 and 40 °C. The comminuted plant is next subjected to a treatment with laser radiation (step c). As source of the laser radiation, a red linear laser diode with a capability of harmonic generation in wavelengths within the range of 150 to 810 nm is preferably employed. The wavelength of the laser radiation is more preferably of 200 to 400 nm and most preferably of 250 nm. The power of the laser radiation is preferably of 1 to 60 watts, more preferably of 10 to 30 watts and most preferably of 20 watts. The spot is preferably of 1 to 6 mm, more preferably of 2 to 5 mm and most preferably of 4 mm of diameter. The comminuted plant is exposed to the laser radiation so that the whole or most of the mixture is irradiated. This is achieved either by displacing manually the laser generator through the comminuted plant, or by passing the comminuted matter on a conveyor belt through a set of several laser generators. Preferably each kilogram of the comminuted matter is treated with the laser radiation for a period of 3 to 10 minutes, more preferably for a period of 5 minutes. Although higher and lower temperatures are not discarded, the temperature at which the comminuted plant is treated with the laser radiation should be between 10 and 40 °C. WO 02/41908 5 PCT/IB00/01947 The laser treated matter is next suspended in water (step d). Any commercial mineral water can be employed in this step. The suspension is effected so that 50 to 300, preferably 100 to 250, grams of the laser treated matter are present per litre of water. Although higher and lower temperatures are not discarded, the temperature at which the comminuted plant is suspended hi water should be between 10 and 40 °C. The suspension is then kept for a period of between 5 to 20 days, preferably of 7 to 15 days, at a temperature of 2 to 10 °C, preferably of 4 to 8°C, so that maceration of the mixture takes place (step e). Finally, after the maceration step, a separation of the liquid phase from the solid phase is effected (step f). The solids can be pressed to facilitate the separation. The separation can be achieved by decantation alone or, preferably, by decantation followed by filtration. The filtration is preferably effected under pressure. Most preferably three consecutive press-filtrations are effected with filters of 5 jam, 1 pm and 0.22 |im. Although higher and lower temperatures are not discarded, the temperature at which the separation is effected should be between 10 and 40 °C. The process of the invention can be applied to any land of plants, both monocotyledomae and dicotyledonae. It can be applied to mixtures of different plants. The whole plant can be submitted to the method of the invention, although leaves and flowers are preferred. Non limitative examples of families of plants to which the method of the invention can be applied are: Asterciceae, Rosaceae, Crucifrae, Labiatae, Equisetaceae, Saxifraganceae, Compositcie, Araliaceae and Umbeliferae. Non limitative examples of species of plants to which the present method can be applied are: Mentha sativa, Pimpinella anisum, Eleutherococcus senticosus, Equinacea angustifolia, Symphytum officinalis and Equisetum arvense. The preferred species to which the method of the invention can be applied are Calendula officinalis, Agrimonia eupatoria, Lepidium latifolium sadLamium album. The aqueous extracts (hydrolates or compositions) obtained by the method of the invention constitute the second aspect of the invention. Thus, compositions resulting from the application of the present method to the non limitative examples: Mentha sativa, Pimpinella anisum, Eleutherococcus senticosus, Equinacea WO 02/41908 6 PCT/IB00/01947 10 angustifolia, Symphytum officinalis and Equiselum arvense fall within the scope of the invention. Of special interest are the aqueous extracts obtained by applying the method of the invention to Calendula officinalis and to the mixture of Agrimonia eupatoria, Lepidium latifolium and Lamium album, which also fall within the scope of the invention. It has been found that some of the aqueous extracts obtained according to the method of the invention find application in medicine. This constitutes, as mentioned above, the third aspect of the invention. The aqueous extract obtained by applying the method of the invention to Calendula oficinalis, and, especially, the aqueous extract obtained by applying the method of the invention to the flowers of this plant presents activity as immune stimulator. This activity being unexpectedly extremely high, as demonstrated in the 15 examples showed below. It is believed, without wanting to be bound to this theory, that the present extract acts stimulating the lymphocyte transformation activity of the lymphocytes T, B and macrophages. The extract obtained by applying the method of the invention to Calendula oficinalis finds, thus, application in the treatment of immune-suppressant diseases. Non limitative examples of these diseases are cancers 20 such as hepatic carcinoma, lung cancer, kidney cancer, colon cancer, breast cancer, prostate cancer or prostatic adenocarcinoma; brain cancers such as astrocytoma and glioblastoma; cervical cancer and bladder cancer; tuberculosis, influenza, common cold, allergies, lupus erythematosus, psoriasis and AIDS. 25 Moreover, the aqueous extract obtained by applying the method of the invention to the mixture of the plants Agrimonia eupatoria, Lepidium latifolium and Lamium album and, especially, the aqueous extract obtained by applying the method of the invention to the mixture of the leaves of the plants Agrimonia eupatoria, Lepidium latifolium and Lamium album unexpectedly presents antiviral activity. 30 More specifically, this extract is especially active against the virus of hepatitis A, B, C, D and E. This activity being extremely high as demonstrated in the examples showed below. Furthermore, this extract also presents a high regulatory activity of the metabolism of transaminases and bilirrubine, as well as activity as stimulator of the hepatocytic regeneration. Therefore, the extract obtained by applying the method 35 of the invention to Agrimonia eupatoria, Lepidium latifolium and Lamium album finds application in the treatment of viral diseases. Non limitative examples of these diseases are hepatitis A, B, C, D and E. This extract finds also application in the IiYI'ELLCCIUAL PROPERTY OFFICE) OF N.Z. ! i i Lv L_ RECEIVED \ WO 02/41908 PCT7IB00/O1947 7 treatment of liver pathologies related to high levels of transaminases and bilirrubine. Non limitative examples of such diseases are hepatitis A, B, C, D and E; hepatic cirrhosis and hepatic carcinoma. According to the studies and experiments made by the inventor, it is believed, without wanting to be bound to this theory, that the laser treatment catalyses reactions between certain compounds of the plants and/or facilitates the extraction of certain compounds of the plants, so that fee presence of these compounds in the extracts results in the unexpected activities referred above. A further aspect of the invention relates to pharmaceutical compositions comprising the aqueous extracts obtained by the present method. The aqueous extracts according to the present invention can be employed either as such or lyophilised for preparing the pharmaceutical compositions. It can be administrated either separately, as aqueous extract or lyophilised, or in the form of pharmaceutical preparations. The drug combination is in the form of a formulation which (1) contains the extract according to the invention alone; (2) contains one or more appropriate binders, carriers and/or further auxiliary materials, and (3) may further contain additional therapeutically active substances. The carrier materials, binders and/or auxiliary materials must be pharmaceutically and pharmacologically tolerable, so that they can be combined with the other components of the formulation or preparation and do not exert adverse effects on the organism treated . The formulations include those which are suitable for oral or parentheral (including subcutaneous, intradermal, intramuscular and intravenous) administration, even though the best route of administration is dependent on the patient's status. Hie formulations can be in the form of single doses. The formulations are prepared according to methods known in the field of pharmacology. The appropriate quantities of active substances suitable for administration may vary as a function of the particularly field therapy. In general, the active substance concentration in a single-dose formulation is 5% to 95% of the total formulation. WO 02/41908 8 PCT/IB00/01947 The invention provided by the application is illustrated by the examples presented hereinbelow. Example 1: Preparation of an aqueous extract of flowers of Calendula oficinalis according to a standard method (comparative example). 500 g. of flowers of Calendula oficinalis are placed in a wash tunnel and subjected to a thorough wash with water at about 28°C. The flowers are next comminuted with a comminuting machine. The resultant 500 g of comminuted matter are next suspended in 2 litres of water at a temperature of about 20 °C. The suspension is then kept for 12 days at a temperature of 4 °C. Finally, the separation of the liquid and the solid phase is effected, first by decantation of the liquid (the solids are pressed to facilitate the separation), and then, by three consecutive press-filtrations with filters of 5, 1 and 0.22 fxm at a temperature of about 20 °C. The process yields approximately 1.7 litres of a solution (aqueous extract) of an ochre colour. Example 2: Preparation of an aqueous extract of flowers of Calendula oficinalis according to the method of the invention. 500 g. of flowers of Calendula oficinalis are placed in a wash tunnel and subjected to a thorough wash with water at about 28°C. The flowers are next comminuted with a comminuting machine. The resultant 500 g of comminuted matter are subjected to a treatment with a red linear laser diode with a capability of harmonic generation in a wavelength of 250 nm, a power of 20 watts and a spot of 4 mm of diameter. The treatment is effected by manually displacing the laser generator through the comminuted matter during 2.5 minutes, so that the whole or most of the mixture is irradiated. The laser treated matter is next suspended in 2 htres of water at a temperature of about 20 °C. The suspension is then kept for 12 days at a temperature of 4 °C. Finally, the separation of the liquid and the solid phase is effected, first by decantation of the liquid (the solids are pressed to facilitate the separation), and then, by three consecutive press-filtrations with filters of 5, 1 and 0.22 |im at a temperature of about 20 °C. The process yields approximately 1.7 htres of a solution (aqueous extract) of an ochre colour. Examples 3-14: The aqueous extracts of the plants listed below were prepared according to the procedures of examples 1 and 2. The parts of the plants which were subjected to the processes mentioned above are indicated in brackets. Equisetum arvense (stems), WO 02/41908 9 PCT/IB0O/O1947 Symphytum officinalis (leaves), Equinacea angustifolia (leaves and flowers), Eleutherococcus senticosus (leaves), Pimpinella anisum (leaves and flowers), and Mentha sativa (leaves). 5 The aqueous extracts obtained in the examples 1-14 were tested in order to establish their activity as immune stimulator by quantifying the lymphocyte transformation activity (LTA). By lymphocyte transformation activity it is meant the fact that the lymphocytes are transformed from a dormant to an active state, which is necessary to fight diseases through an immunological mechanism, or to restore the 10 immune system, which might be weakened by different factors. These tests were performed in vitro by adding the extracts to lymphocytes isolated from mice according to the literature reference Max, W. et al, Journal of Natural Products, vol. 54, no. 6, pp. 1531-1542 (1991). The incorporation of thymidine, which means replication of DNA, was monitored. This incorporation is indicative both of an 15 increase in lymphocyte number and an increase in lymphocyte activity. The results are summarised in table 1. Table! Examples Plant Increase LTA (%) (Standard method) Increase LTA (%) (Method including laser treatment) 1/2 Calendula oficinalis + 277 + 1204 3/4 Equisetum arvense + 26 + 123 5/6 Symphytum officinalis + 43 + 211 7/8 Echinacea angustifolia + 98 + 270 9/10 Eleutherococcus senticosus + 106 + 280 11/12 Pimpinella anisum + 11 + 26 13/14 Mentha sativa + 12 + 28 20 From the results showed in table 1, it is clear how the aqueous extracts prepared according to the method of the invention present higher increases in the lymphocyte transformation activity values than the aqueous extracts obtained according to the standard methods, i.e., extraction without laser treatment. 25 Furthermore, it is really surprising the extremely high increase in the lymphocyte WO 02/41908 10 PCT/IB00/01947 transfonnation activity value showed by the extract of Calendula officinalis obtained according to the method of the invention Example 15: Preparation of an aqueous extract of the leaves of the plants Agrimonia eupatoria, Lepidium latifolium and Lamium album according to a standard method (comparative example). 250 g. of leaves of Agrimonia eupatoria, 250 g. of leaves of Lepidium latifolium and 250 g. of leaves of Lamium album, are placed in a wash tunnel and subjected to a thorough wash with water at about 28°C. The leaves were next comminuted with a comminuting machine. The resultant 750 g of comminuted matter are next suspended in 3 litres of water at a temperature of about 20 °C. The suspension is then kept for 12 days at a temperature of 4 °C. Finally, the separation of the liquid and the solid phase is effected, first by decantation of the liquid (the solids are pressed to facilitate the separation), and then, by three consecutive press-filtrations with filters of 5, 1 and 0.22 fim at a temperature of about 20 °C. The process yields approximately 2.7 litres of a solution (aqueous extract) of an dark green colour. Example 16: Preparation of an aqueous extract of the leaves of the plants Agrimonia eupatoria, Lepidium latifolium and Lamium album according to the method of the invention. 250 g. of leaves of Agrimonia eupatoria, 250 g. of leaves of Lepidium latifolium and 250 g. of leaves of Lamium album, axe placed in a wash tunnel and subjected to a thorough wash with water at about 28°C. The leaves were next commhiuted with a comminuting machine. The resultant 750 g of comminuted matter are submitted to a treatment with a red linear laser diode with a capability of harmonic generation in a wavelength of 250 nm, a power of 20 watts and a spot of 4 mm of diameter. The treatment is effected by manually displacing the laser generator through the comminuted matter during 4 minutes, so that the whole or most of the mixture is irradiated. The laser treated matter is next suspended in 3 litres of water at a temperature of about 20 °C. The suspension is then kept for 12 days at a temperature of 4 °C. Finally, the separation of the liquid and the solid phase is effected, first by decantation of the liquid (the solids are pressed to facilitate the separation), and then, by three consecutive press-filtrations with filters of 5, 1 and 0.22 jj.m at a temperature of about 20 °C. The process yields approximately 2.7 htres of a solution (aqueous extract) of ail dark green colour. </div>

Claims

1. WO 2. /41908 11 PCT/IBOO/01947 The aqueous extracts obtained hi the examples 15 and 16 were employed in clinical tests on 28 human beings suffering from different hepatic diseases and, consequently, presenting high values of transaminases (GOT, GPT, GGT), bilirrubine and high viral charges. Those patients treated with the extract obtained in example 15 presented, after three months of treatment, a slight decrease in the transaminases and no variation of the viral charges. However, those patients treated with the extract obtained in example 16 surprisingly presented, after two months of treatment, a symptomatic recovery, normal values of transaminases and bilirrubine, as well as an important decrease of the viral charges, even in the case of hepatitis C of strain 1-B and mutated virus. 12 WHAT WE CLAIM IS: 1. Method for preparing aqueous extracts of plants which comprises the following a) Decontamination of the plant b) Comminuting the plant. c) Treatment of the comminuted plant with a laser radiation. d) Suspension of the mixture obtained in step c) in water. e) Maceration of the suspension obtained in step d). f) Separation of the resulting liquid.

2. Method according to claim 1, wherein the method is applied to a plant unit, to several plant units of the same species or to several plant units of different species.

3. Method according to claim 1 or claim 2, wherein the method is applied to a part or parts of a plant such as stems, leaves or flowers.

4. Method according to any one of the preceding claims, wherein the decontamination is effected with water.

5. Method according to claim 4, wherein the decontamination is effected in a washing tunnel.

6. Method according to any one of the preceding claims, wherein the comminuting is effected manually or with a comminuting machinery.

7. Method according to any one of the preceding claims, wherein the treatment with the laser radiation is effected with a red linear laser diode with a capability of harmonic generation in wavelengths within the range of 150 to 810 nm, a power of 1 to 60 watts and a spot of 1 to 6 mm of diameter.

8. Method according to claim 7, wherein the wavelength is within the range of 200 to 400 nm, a power of 20 watts and a spot of 4 mm of diameter. 13 9. Method according to claim 8, wherein the wavelength is 250 nm. 10. Method according to any one of the preceding claims, wherein each kilogram of the comminuted matter is treated with the laser radiation for a period of 3 to 10 minutes. 11. Method according to claim 10, wherein each kilogram of the comminuted matter is treated with the laser radiation for a period of 5 minutes. 12. Method according to any one of the preceding claims, wherein the suspension is effected so that 50 to 300 grams of the laser treated matter are present per litre of water. 13. Method according to claim 12, wherein the suspension is effected so that 100 to 250 grams of the laser treated matter are present per litre of water. 14. Method according to any one of the preceding claims, wherein the suspension is kept for a period of between 5 to 20 days at a temperature of 2 to 10°C, so that maceration takes place. 15. Method according to claim 14, wherein the suspension is kept for period of between 7 to 15 days. 16. Method according to claim 14 or claim 15, wherein the temperature is 4 to 8°C. 17. Method according to any one of the preceding claims, wherein the liquid phase is separated from the solid phase by effecting a decantation followed by a filtration consisting of three consecutive press-filtrations with filters of 5,1 and 0.22 jam. 18. Method according to any one of the preceding claims, wherein the method is applied to monocotyledonae or dicotyledonae plants. | INTELLECTU-Tr^crtYY Os-RCEI I OF N.Z. I J 14 OCT 2334 | i •-HC£iV:::0 19. 20, 21. 22. 23, 24, 25, 26 27, 28 14 Method according to any one of the preceding claims, wherein the plant belongs to the following families: Asteraceae, Rosaceae, Crucifrae, Labiatae, Equiesetaceae, Saxifragancaea, Compositae, Araliceae and Umbeliferae. Method according to any one of the preceding claims, wherein the plant is chosen from the following species: Mentha sativa, Pimpinella anisum, Eleutherococcus senticosus, Equinacea angustifolia, Symphytum officinalis, Equisetum arvense, Calendula officinalis, Agrimonia eupatoria, Lepidium latifolium and Laminium album. Method according to any one of the preceding claims, wherein the plant is Calendula officinalis. Method according to claim 21, wherein the method is applied to the flowers of Calendula officinalis. Method according to any one of claims 1 to 20, wherein the method is applied to a mixture of Agrimonia eupatoria, Lepidium latifolium and Lamium album. Method according to claim 23, wherein the method is applied to the leaves of Agrimonia eupatoria, Lepidium latifolium and Lamium album. Aqueous extracts obtained according to any of the methods defined in claims 1 to 20. Lyophilised aqueous extracted according to claim 25. Aqueous extracts obtained according to any of the methods defined in claims 21 and 22. Lyophilised aqueous extracts according to claim 27. 15 29. 30. 31. 32. 36. 37. 38. Aqueous extracts obtained according to any of the methods defined in claims 23 and 24. Lyophilised aqueous extracts according to claim 29. Aqueous extracts or lyophilised aqueous extracts according to any one of claims 25 to 30 for use as therapeutic agents. Aqueous extracts or lyophilised aqueous extracts according to claim 27 or claim 28 for use in the treatment of immune suppressant diseases. Aqueous extracts or lyophilised aqueous extracts according to claim 27 or claim 28 for use in the treatment of cancer, tuberculosis, influenza, common cold, allergies, lupus erythematosus, psoriasis and AIDS. Aqueous extracts of lyophilised aqueous extracts according to claim 27 or claim 28 for use in the treatment of hepatic carcinoma, lung cancer, kidney cancer, colon cancer, breast cancer, prostate cancer or prostatic adenocarcinoma; brain cancers such as strocytoma and glioblastoma; cervical cancer and bladder cancer. Aqueous extracts or lyophilised aqueous extracts according to claim 29 or claim 30 for use in the treatment of viral diseases. Aqueous extracts or lyophilised aqueous extracts according to claim 29 or claim 30 for use as stimulators for the regeneration of hepatocytes. Aqueous extracts according to claim 29 or claim 30 for use in the treatment of pathologies related to high levels of transaminases and bilirrubine. Aqueous extracts according to claim 29 or claim 30 for use in the treatment of hepatitis A, B, C, D and E; hepatic cirrhosis and hepatic carcinoma. 16 39. Use of the aqueous extracts or lyophilised aqueous extracts according to any one of claims 25 to 30 for the manufacture of a medicament. 40. Use of the aqueous extracts or lyophilised aqueous extracts according to any one of claims 27 to 30 for the manufacture of a medicament for treating immune suppressant diseases. 41. Use according to claim 40 for the manufacture of a medicament for treating cancer, tuberculosis, influenza, common cold, allergies, lupus erythematosus, psoriasis and AIDS. 42. Use according to claim 41 for the manufacture of a medicament for treating hepatic carcinoma, lung cancer, kidney cancer, colon cancer, breast cancer, prostate cancer or prostatic adenocarcinoma; brain cancers such as strocytoma and glioblastoma; cervical cancer and bladder cancer. 43. Use of the aqueous extracts or lyophilised extracts according to claim 29 or claim 30 for the manufacture of a medicament for treating viral diseases. 44. Use according to claim 43 for the manufacture of a medicament for the treatment of hepatitis A, B, C, D and E. 45. Use of the aqueous extracts or lyophilised aqueous extracts according to claim 29 or claim 30 for the manufacture of a medicament for the treatment of pathologies related to high levels of transaminases and bilirrubine. 46. Use according to claim 45 for the manufacture of a medicament for the treatment of hepatic cirrhosis and hepatic carcinoma. 47. Use of the aqueous extracts or lyophilised aqueous extracts according to claim 29 or claim 30 for the manufacture of a medicament for stimulating hepatocytic regeneration. f 4 oc: i 48. 49. 50. 51. 52. 53, 54, 55, 56, 57 58 59 60 17 A pharmaceutical preparation comprising an aqueous extract according to any one of claims 25 to 30. The pharmaceutical preparation according to claim 48 further including a pharmaceutically acceptable vehicle. The pharmaceutical preparation of claim 47 or claim 48 further including at least one other pharmaceutically active compound. A method according to claim 1 substantially as herein described or exemplified. An aqueous extract according to claim 25 substantially as herein described or exemplified. An aqueous extract according to claim 27 substantially as herein described or exemplified. An aqueous extract according to claim 29 substantially as herein described or exemplified. A use according to claim 39 substantially as herein described or exemplified. A use according to claim 40 substantially as herein described or exemplified. A use according to claim 43 substantially as herein described or exemplified. A use according to claim 45 substantially as herein described or exemplified. A use according to claim 47 substantially as herein described or exemplified. A pharmaceutical preparation according to claim 48 substantially as herein described or exemplified.